Single-cell RNA sequencing of healthy and dystrophic murine skeletal muscle

Fibrosis and fat replacement in the skeletal muscle is a major complication that leads to a loss of mobility in chronic muscle disorders, such as muscular dystrophy. However, our current knowledge on the in vivo properties of adipogenic stem and precursor cells remains unclear, mainly due to the high cell heterogeneity in skeletal muscles. For this purpose, we used single-cell RNA-sequencing to decomplexify interstitial cell populations in healthy and dystrophic skeletal muscles. We identified a CD142 (F3) positive cell subpopulation in mice and humans that is responsible for the inhibition of adipogenesis. Furthermore, we found a completely altered composition of interstitial cells in muscular dystrophy, with a near absence of the CD142-positive cells. The novel discovery of these adipo-regulatory cells in the skeletal muscle aids our understanding regarding the aberrant fat deposition in muscular dystrophy, paving the way for treatments that potentially sustain ambulation in patients with muscular dystrophy. Hindlimb muscles of wild-type (C57BL6, n=1) and dystrophic (Sgcb-null, n=1) mice were digested and sorted as CD45-negative and CD31-negative single-cells in 96w plates. A total of 384 cells were processed into a single-cell library with SMART-seq2.